Linkage of the scrapie-associated fibril protein (PrP) gene and Sinc using congenic mice and restriction fragment length polymorphism analysis.

Linkage of the scrapie-associated fibril protein (PrP) gene and Sinc using congenic mice and restriction fragment length polymorphism analysis.
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使用同源小鼠和限制性片段长度多态性分析分析痒病相关原纤维蛋白 (PrP) 基因和 Sinc。

DOI:
10.1099/0022-1317-68-10-2711
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发表时间:
1987
期刊:
The Journal of general virology
影响因子:
--
通讯作者:
A. Dickinson
A. Dickinson
中科院分区:
--
文献类型:
--
作者:
N. Hunter;James Hope;I. Mcconnell;A. Dickinson

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Sinc具有两个等位基因p7和s7,是决定所有羊瘙痒病菌株在小鼠中潜伏期的主要基因。羊瘙痒病相关原纤维的主要蛋白(PrP)是由细胞基因编码的,我们使用了仓鼠PrP mRNA的cDNA拷贝进行限制性片段长度多态性(RFLP)分析不同的近交系小鼠品系,包括VM(Sincp7)和VM(Sincs7)同源小鼠。在VM(Sincp7)小鼠中,5.5kb的XbaI片段与PrP cDNA序列杂交,而VM(Sincs7)同源小鼠具有3.8kb的XbaI片段。VM × VM(Sincs7)同系F1小鼠同时具有5.5kb和3.8kb片段。Sincs7供体小鼠品系C57BL具有3.8kb片段,表明Sinc基因和编码PrP的基因是连锁的,甚至可能是相同的基因。其他Sincp7近交系小鼠(IM和MB)有5.5 kb的片段,但也有一些Sincs7菌株(RIII和VL),这意味着XbaI位点多态性在功能上不涉及两个Sinc等位基因之间的差异。我们已经将多态性XbaI位点定位到PrP基因的3'侧翼区。TaqI和HhaI也被发现在所研究的近交系小鼠品系中显示多态性。HhaI的明显RFLP可能是甲基化差异而不是序列差异的结果。
Sinc, with two alleles p7 and s7, is the major gene determining the incubation period of all strains of scrapie in mice. The major protein (PrP) of scrapie-associated fibrils is encoded by a cellular gene and we have used a cDNA copy of the hamster PrP mRNA to carry out restriction fragment length polymorphism (RFLP) analysis of different inbred mouse strains including VM(Sincp7) and VM(Sincs7) congenic mice. In VM(Sincp7) mice, a 5.5 kb XbaI fragment hybridized to the PrP cDNA sequence whereas VM(Sincs7) congenic mice had a 3.8 kb XbaI fragment. The VM X VM(Sincs7) congenic F1 mice had both the 5.5 kb and the 3.8 kb fragments. The Sincs7 donor mouse strain, C57BL, had the 3.8 kb fragment suggesting that the Sinc gene and the gene coding for PrP are linked, and could even be the same gene. Other Sincp7 inbred mice (IM and MB) had the 5.5 kb fragment but so too did some Sincs7 strains (RIII and VL), implying that the XbaI site polymorphism is not functionally involved in the difference between the two Sinc alleles. We have mapped the polymorphic XbaI site to the 3' flanking region of the PrP gene. TaqI and HhaI were also found to show polymorphisms in the inbred mouse strains studied. The apparent RFLP with HhaI may be a result of differences in methylation rather than in sequence.
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